Broad-Host-Range Plasmids for Constitutive and Inducible Gene Expression in the Absence of Antibiotic Selection

Source: Erdogan, F.,et al., Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation. J. Vis. Exp. (2017)This video demonstrates the procedure for transferring an F plasmid from donor to recipient E. coli cells using a rescue plasmid and selecting transconjugants through dual antibiotic resistance. The process includes mixing bacterial cultures, incubation without shaking, disrupting mating pairs, and plating on selective media to confirm...

Here, we present a framework to relate broad-range dietary restriction to gene expression and lifespan. We describe protocols for broad-range dietary restriction and for quantitative imaging of gene expression under this paradigm. We further outline computational analyses to reveal underlying information processing features of the genetic circuits involved in...

Overview

This is a method to generate “scarless” recombinant vaccinia viruses using host-range selection and visual identification of recombinant...

Gene expression in prokaryotes is governed by constitutive and regulated systems, allowing cells to balance the production of essential proteins with adaptive responses to environmental changes.Constitutive Gene ExpressionConstitutive, or housekeeping, genes are continuously expressed as they encode proteins vital for fundamental cellular processes. These include enzymes for glycolysis, ribosomal components for protein synthesis, and proteins involved in DNA replication. Their constant...

Our experimental approach provides a strategy to follow plasmid abundance and antibiotic resistance over time in bacterial...